Quantitative characterization of single cells by use of immunocytochemistry combined with multiplex LA-ICP-MS

Abstract

Actual research demonstrates that LA-ICP-MS is capable of being used as an imaging tool with cellular resolution. The aim of this investigation was the method development for LA-ICP-MS to extend the versatility to quantitative and multiplexing imaging of single eukaryotic cells. For visualization of individual cells selected, lanthanide-labeled antibodies were optimized for immuno-imaging of single cells with LA-ICP-MS. The molar content of the artificial introduced labels per cell was quantified using self-made nitrocellulose-coated slides for matrix-matched calibration and calculated amounts were in the range of 3.1 to 17.8 atmol per cell. Furthermore, the quantification strategy allows a conversion of 2D intensity profiles based on counts per second (cps) to quantitative 2D profiles representing the molar amount of the artificial introduced elemental probes per pixel for each individual cell. Graphical abstract ᅟ.

Quantification of silver nanoparticles taken up by single cells using inductively coupled plasma mass spectrometry in the single cell measurement mode

“…In both studies the cell system was not synchronized, which means that cells are in different activity cycles once the experiment is started and thus have different uptake rates and even different uptake times which might deviate from the incubation time signicantly. 16 The AgNP uptake by both THP-1 cell types (monocytes and partially differentiated macrophages) is comparable at least within the uncertainty ranges. Even so, the AgNP uptake is increased for THP-1 partially differentiated macrophage cells (Fig.…”

Isotopically Enriched Tracers and Inductively Coupled Plasma Mass Spectrometry Methodologies to Study Zinc Supplementation in Single-Cells of Retinal Pigment Epithelium in Vitro

“…For characterization of single-cells by LA-ICP-MS imaging, the use of metal staining reagents for cell localization was previously reported. 20,24 In our experiments, Fig. 3a shows the microscope image of the selected area, distinguishing the cells from the glass substrate of the chamber-slide.…”

“…Interesting advances related to reduced spot sizes, ultra-fast wash-out ablation cells and data analysis software have been reported for LA-ICP-MS, allowing to obtain images from cells. 13,[18][19][20][21][22] For example, Pisonero et al 13 reported qualitative images of Au and Cd elemental distribution within mouse embryonic fibroblast cells and human cervical carcinoma cells, incubated with Au nanoparticles and Cd-based quantum dots, respectively. Moreover, different exogenous labels (e.g.…”

“…single metal chelates such as DOTA lanthanide complexes and polymeric labels such as MAXPAR®) were investigated for imaging of proteins within single cells. 14,20,23,24 Furthermore, some strategies based on standards spotted onto nitrocellulose membranes or microarray gelatin standards were used to obtain not only elemental qualitative distributions but also quantitative information from cell cultures. 14,25 The aim of our investigation was to evaluate the Zn absorption in cultured HRPEvs cells after its supplementation with isotopically-enriched tracers ( t68 Zn and t70 Zn).…”

Quantitative mapping of specific proteins in biological tissues by laser ablation–ICP-MS using exogenous labels: aspects to be considered

“…Printing of metal spiked inks onto tissue sections has been also proposed for internal standardization [40]. Moreover, Mueller et al [41] presented an interesting approach, using lanthanide-labelled antibodies, for quantitative immunoimaging of single eukaryotic cells; the calculation of an exact labelling degree as well as the correlation factor between amount of lanthanide per cell and amount of antibodyantigen complex were reported as critical aspects for further optimization.…”